Lysis Buffer

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Joery De Kock - One of the best experts on this subject based on the ideXlab platform.

  • proliferative and phenotypical characteristics of human adipose tissue derived stem cells comparison of ficoll gradient centrifugation and red blood cell Lysis Buffer treatment purification methods
    Cytotherapy, 2014
    Co-Authors: Mehdi Najar, Karolien Buyl, Tamara Vanhaecke, Vera Rogiers, Robim Marcelino Rodrigues, Steven Branson, Laurence Lagneaux, Joery De Kock
    Abstract:

    Abstract Background aims Adult human subcutaneous adipose tissue harbors a multipotent stem cell population, the so-called human adipose tissue–derived mesenchymal stromal cells (AT-MSCs). These cells are able to differentiate in vitro into various cell types and possess immunomodulatory features. Yet procedures to obtain AT-MSCs can vary significantly. The two most extensively used AT-MSC purification techniques are (i) density gradient centrifugation using Ficoll and (ii) red blood cell (RBC) Lysis Buffer treatment of the stromal vascular fraction. In the context of potential clinical cell therapy, the stem cell yield after purification and upon consecutive passages, as well as the purity of the obtained cell population, are of utmost importance. Methods We investigated the expansion capacity and purity of AT-MSCs purified by both procedures immediately after isolation and upon consecutive passages. We also investigated possible purification-dependent differences in their expression of immune-inhibitory factors and cell adhesion molecules. Results We found that RBC Lysis Buffer treatment is a more robust and easier method to purify AT-MSCs than density gradient fractionation. However, the resulting AT-MSC-RBC population contains a significantly higher number of CD34 + cells, particularly during the first passages after plating. From passage 4 onward, no significant differences could be observed between both populations with respect to the immunophenotype, expansion capacity and expression of immune inhibitory factors and cell adhesion molecules. Conclusions Our data show that RBC Lysis Buffer treatment may be a good alternative to density fractionation, providing a faster, more robust and easier method to purify AT-MSCs with biologically preserved characteristics.

Mehdi Najar - One of the best experts on this subject based on the ideXlab platform.

  • proliferative and phenotypical characteristics of human adipose tissue derived stem cells comparison of ficoll gradient centrifugation and red blood cell Lysis Buffer treatment purification methods
    Cytotherapy, 2014
    Co-Authors: Mehdi Najar, Karolien Buyl, Tamara Vanhaecke, Vera Rogiers, Robim Marcelino Rodrigues, Steven Branson, Laurence Lagneaux, Joery De Kock
    Abstract:

    Abstract Background aims Adult human subcutaneous adipose tissue harbors a multipotent stem cell population, the so-called human adipose tissue–derived mesenchymal stromal cells (AT-MSCs). These cells are able to differentiate in vitro into various cell types and possess immunomodulatory features. Yet procedures to obtain AT-MSCs can vary significantly. The two most extensively used AT-MSC purification techniques are (i) density gradient centrifugation using Ficoll and (ii) red blood cell (RBC) Lysis Buffer treatment of the stromal vascular fraction. In the context of potential clinical cell therapy, the stem cell yield after purification and upon consecutive passages, as well as the purity of the obtained cell population, are of utmost importance. Methods We investigated the expansion capacity and purity of AT-MSCs purified by both procedures immediately after isolation and upon consecutive passages. We also investigated possible purification-dependent differences in their expression of immune-inhibitory factors and cell adhesion molecules. Results We found that RBC Lysis Buffer treatment is a more robust and easier method to purify AT-MSCs than density gradient fractionation. However, the resulting AT-MSC-RBC population contains a significantly higher number of CD34 + cells, particularly during the first passages after plating. From passage 4 onward, no significant differences could be observed between both populations with respect to the immunophenotype, expansion capacity and expression of immune inhibitory factors and cell adhesion molecules. Conclusions Our data show that RBC Lysis Buffer treatment may be a good alternative to density fractionation, providing a faster, more robust and easier method to purify AT-MSCs with biologically preserved characteristics.

Tamara Vanhaecke - One of the best experts on this subject based on the ideXlab platform.

  • proliferative and phenotypical characteristics of human adipose tissue derived stem cells comparison of ficoll gradient centrifugation and red blood cell Lysis Buffer treatment purification methods
    Cytotherapy, 2014
    Co-Authors: Mehdi Najar, Karolien Buyl, Tamara Vanhaecke, Vera Rogiers, Robim Marcelino Rodrigues, Steven Branson, Laurence Lagneaux, Joery De Kock
    Abstract:

    Abstract Background aims Adult human subcutaneous adipose tissue harbors a multipotent stem cell population, the so-called human adipose tissue–derived mesenchymal stromal cells (AT-MSCs). These cells are able to differentiate in vitro into various cell types and possess immunomodulatory features. Yet procedures to obtain AT-MSCs can vary significantly. The two most extensively used AT-MSC purification techniques are (i) density gradient centrifugation using Ficoll and (ii) red blood cell (RBC) Lysis Buffer treatment of the stromal vascular fraction. In the context of potential clinical cell therapy, the stem cell yield after purification and upon consecutive passages, as well as the purity of the obtained cell population, are of utmost importance. Methods We investigated the expansion capacity and purity of AT-MSCs purified by both procedures immediately after isolation and upon consecutive passages. We also investigated possible purification-dependent differences in their expression of immune-inhibitory factors and cell adhesion molecules. Results We found that RBC Lysis Buffer treatment is a more robust and easier method to purify AT-MSCs than density gradient fractionation. However, the resulting AT-MSC-RBC population contains a significantly higher number of CD34 + cells, particularly during the first passages after plating. From passage 4 onward, no significant differences could be observed between both populations with respect to the immunophenotype, expansion capacity and expression of immune inhibitory factors and cell adhesion molecules. Conclusions Our data show that RBC Lysis Buffer treatment may be a good alternative to density fractionation, providing a faster, more robust and easier method to purify AT-MSCs with biologically preserved characteristics.

Vera Rogiers - One of the best experts on this subject based on the ideXlab platform.

  • proliferative and phenotypical characteristics of human adipose tissue derived stem cells comparison of ficoll gradient centrifugation and red blood cell Lysis Buffer treatment purification methods
    Cytotherapy, 2014
    Co-Authors: Mehdi Najar, Karolien Buyl, Tamara Vanhaecke, Vera Rogiers, Robim Marcelino Rodrigues, Steven Branson, Laurence Lagneaux, Joery De Kock
    Abstract:

    Abstract Background aims Adult human subcutaneous adipose tissue harbors a multipotent stem cell population, the so-called human adipose tissue–derived mesenchymal stromal cells (AT-MSCs). These cells are able to differentiate in vitro into various cell types and possess immunomodulatory features. Yet procedures to obtain AT-MSCs can vary significantly. The two most extensively used AT-MSC purification techniques are (i) density gradient centrifugation using Ficoll and (ii) red blood cell (RBC) Lysis Buffer treatment of the stromal vascular fraction. In the context of potential clinical cell therapy, the stem cell yield after purification and upon consecutive passages, as well as the purity of the obtained cell population, are of utmost importance. Methods We investigated the expansion capacity and purity of AT-MSCs purified by both procedures immediately after isolation and upon consecutive passages. We also investigated possible purification-dependent differences in their expression of immune-inhibitory factors and cell adhesion molecules. Results We found that RBC Lysis Buffer treatment is a more robust and easier method to purify AT-MSCs than density gradient fractionation. However, the resulting AT-MSC-RBC population contains a significantly higher number of CD34 + cells, particularly during the first passages after plating. From passage 4 onward, no significant differences could be observed between both populations with respect to the immunophenotype, expansion capacity and expression of immune inhibitory factors and cell adhesion molecules. Conclusions Our data show that RBC Lysis Buffer treatment may be a good alternative to density fractionation, providing a faster, more robust and easier method to purify AT-MSCs with biologically preserved characteristics.

Laurence Lagneaux - One of the best experts on this subject based on the ideXlab platform.

  • proliferative and phenotypical characteristics of human adipose tissue derived stem cells comparison of ficoll gradient centrifugation and red blood cell Lysis Buffer treatment purification methods
    Cytotherapy, 2014
    Co-Authors: Mehdi Najar, Karolien Buyl, Tamara Vanhaecke, Vera Rogiers, Robim Marcelino Rodrigues, Steven Branson, Laurence Lagneaux, Joery De Kock
    Abstract:

    Abstract Background aims Adult human subcutaneous adipose tissue harbors a multipotent stem cell population, the so-called human adipose tissue–derived mesenchymal stromal cells (AT-MSCs). These cells are able to differentiate in vitro into various cell types and possess immunomodulatory features. Yet procedures to obtain AT-MSCs can vary significantly. The two most extensively used AT-MSC purification techniques are (i) density gradient centrifugation using Ficoll and (ii) red blood cell (RBC) Lysis Buffer treatment of the stromal vascular fraction. In the context of potential clinical cell therapy, the stem cell yield after purification and upon consecutive passages, as well as the purity of the obtained cell population, are of utmost importance. Methods We investigated the expansion capacity and purity of AT-MSCs purified by both procedures immediately after isolation and upon consecutive passages. We also investigated possible purification-dependent differences in their expression of immune-inhibitory factors and cell adhesion molecules. Results We found that RBC Lysis Buffer treatment is a more robust and easier method to purify AT-MSCs than density gradient fractionation. However, the resulting AT-MSC-RBC population contains a significantly higher number of CD34 + cells, particularly during the first passages after plating. From passage 4 onward, no significant differences could be observed between both populations with respect to the immunophenotype, expansion capacity and expression of immune inhibitory factors and cell adhesion molecules. Conclusions Our data show that RBC Lysis Buffer treatment may be a good alternative to density fractionation, providing a faster, more robust and easier method to purify AT-MSCs with biologically preserved characteristics.